高产褐藻酸酶菌株的筛选、发酵条件优化和褐藻酸酶酶学性质的初步研究-细胞生物学专业论文.docxVIP

Study on Screening and Selecting an Effectively Alginase-producing Strain，its Optimal Cultivation and Characteristics ofAlginase Abstract With the purposes of screening and selecting all effectively alginase-producing strain，studying the biological effect and characteristics ofalginase and using alginase to produce alginate oligosaccharides．several strains have been isolated with alginase activity from the decaying parts of Laminaria japonica．Among these stralii5，the strain s4 showed the highest alginaso activity during the second screening and demonstrates a good stability,so it was chosen to investigate the fermentation condition and some properties ofalginase produced by strain s4． There are many factors that influence the alginase production．It was found that the strain can not produce alginase without sodium alginate．The different carbon SoIIrces tests also showed that the alginase of strain s4 Was an induced enzylne．In one—factor-at-a-time methodology experiment，the optimal conditions of strain s4 to produce algtlia∞were studied．The optimal fermentation medium of strain s4 contained sodium alginate 1．2％；NI-hCI O．9％；NaCI 1．5％initial pH 7．5．The optimal temperature and time of fermentation conditions for s4 to produce alginase were 25C and 48h，respectively．Under the condition,the strain s4 would produce the highest amount ofalgmase． Alginase produced by strain s4 Was all inducible and secreting extracellar Calzyme．The aMnase produced by the strain s4 Was purified to an apparent  homogeneity identified by SDS·PAGE through ammonium sulfate precipitation (20．80％saturation)，CM—Sepharose Fast Flow chromatography,and Sephacryl S-1 00 gd filtration．Molecular weights of the eIlzymes were estimated to be 29 kDa． Alginase activity WaS stable in the pH range of 6．0-9．0 and under 45C，and the optimal pH value and temperature ofaiginase were 7．5 and 40C．The enzyme activity WaS significantly inhibited by H92+and slightly activated by M92+，Na*and K+at ion concentration being 5